Detection and Identification of Gastrointestinal Pathogens in Clinical Samples: Comparison Between two Laboratory Developed Multiplex Real-Time PCR Platforms

Graduation Date


Document Type

Master's Thesis

Degree Name

Master of Science

Department or Program

Clinical Laboratory Sciences

Department or Program Chair

Mary Sevigny, PhD

First Reader

LaRonda S. Frazier

Second Reader

Maria C. DeSousa, JD, MPA, CLS


Infectious gastroenteritis is caused by a wide range of enteric bacteria pathogens including Salmonella, Shigella, Campylobacter jejuni, and E. coli 0157. Conventional methods used in the detection of such pathogens including, culture, microscopy, biochemical identification, and serological testing, are labor intensive, time-consuming, and lack specificity and sensitivity. In recent years, multiplex molecular testing has made its way into the clinical laboratory for detection and identification of enteric pathogens directly from clinical stool samples. Multiplex molecular assays are able to identify several gastrointestinal pathogens in as little as a few hours. This paper compares the test methodology, workflow, turnaround time, and cost-effectiveness of two laboratory developed multiplex PCR assays referred to as Platform A and Platform B used in the detection and identification of six enteric pathogens. This analysis comparing the current (Platform A) to newly proposed (Platform B), found the implementation of platform B into our current workflow to be advantageous. The implementation of platform B will significantly reduce the analysis to reporting turnaround time by 5 hours. A reduction in turnaround time will help clinicians diagnose gastroenteritis in a more accurate and timely manner. Reducing delay of treatment, inconsistent antimicrobial therapy, and/or expensive medical care.

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